Purification of a New Elicitin from Phytopthora cambivora KACC40160

Phytophthora cambivora KACC 40160으로부터 새로운 elicitin의 분리

  • Yoon, Sang-Hong (Metabolic Engineering Division, National Institute of Agricultural Biotechnology Rural Development Administration) ;
  • Bae, Shin-Chul (Molecular Physiology Division, National Institute of Agricultural Biotechnology Rural Development Administration) ;
  • Park, In-Cheol (Molecular Physiology Division, National Institute of Agricultural Biotechnology Rural Development Administration) ;
  • Koo, Bon-Sung (Metabolic Engineering Division, National Institute of Agricultural Biotechnology Rural Development Administration) ;
  • Kim, Young-Hwan (Metabolic Engineering Division, National Institute of Agricultural Biotechnology Rural Development Administration) ;
  • Yeo, Yun-Soo (Metabolic Engineering Division, National Institute of Agricultural Biotechnology Rural Development Administration)
  • 윤상홍 (농업생명공학연구원 신기능소재개발과) ;
  • 배신철 (농업생명공학연구원 분자생리과) ;
  • 박인철 (농업생명공학연구원 신기능소재개발과) ;
  • 구본성 (농업생명공학연구원 신기능소재개발과) ;
  • 김용환 (농업생명공학연구원 신기능소재개발과) ;
  • 여윤수 (농업생명공학연구원 신기능소재개발과)
  • Published : 2003.05.31

Abstract

Elicitins, proteinaceous elicitors secreted from Oomycetes fungi (Phytophthora spp. and Pythium spp.), have been known as inducer of hypersensitive response (HR) in incompatible interactions between plant and pathogens. Five elicitins among many Korean Phytophthora species caused the reactions of distal HR in radish, chinese cabbage and some hot pepper cultivars, but not in cucumber and tomato. Because the isolation of elicitin from Phytophthora cambivora hasn't been reported yet, we have purified a cambivorein, a new member of the elicitin family, from the culture filtrate of Phytophtilora cambivora (KACC 40160) by using FPLC (Fast Protein Liquid Chromatography, AKTA) with sepharose S and Sephacryl HR columns. We confirmed that it induces necrosis activities in some hot pepper cultivars and its molecular weight is about 10 KDa by Tricine-SDS-PAGE. Comparison of amino acid sequences of its N-terminal ends also informed the identification of Iysine at the 13th position, which is characteristic of a kind of basic elicitin isoform $({\beta}-isoform)$. It Also showed that our elicitin is not identical with N-terminal sequences of many elicitins reported from Phytophthora spp..

난균류에 속하는 Phytophthora spp.와 pythium spp.에서 분비되는 단백질인 elicitin은 식물과 병원균과의 비친화적 관계에서 과민감반응을 유도하는 인자로 알려져 왔다. 국내에서 수집된 5종의 Phytophthora spp로부터 분리된 elicitin들은 무, 배추, 고추에서 과민감 반응을 유도 하였으나 오이, 토마토에서는 볼 수 없었다. 특히 지금까지 보고가 되지않은 P. cambivora KACC 40160균주의 배양액으로부터 ion exchange와 gel filtration을 이용하여 고추에 과민감반응을 유도하며 분자량이 10kDa인 새로운 elicitin을 순수분리 하여 cambivorein이라 명명 하였다. 분리된 cambivorein의 N-말단 아미노산 서열분석 결과 ${\beta}-isoform$의 특징인 13번째 아미노산이 Iysine을 가지고 있었으며 기존의 다른 종류의 elicitin과는 다른 아미노산으로 구성되어 있었다.

Keywords

References

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